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13 February 2013

Detection of Pneumocystis in the nasal swabs of immune-suppressed rats by use of PCR and microscopy

Hüseyin CanABCDE, Ayşe CanerABCDG, Mert DöşkayaACDEG, Aysu DeğirmenciBC, Sabire KaraçalıAD, Ceylan PolatBC, Yüksel GürüzADG, Ahmet ÜnerADG

DOI: 10.12659/MSMBR.883777

Med Sci Monit Basic Res 2013; 19:62-67

Abstract

BACKGROUND: Detection of Pneumocystis jiroveci colonization in lungs or oral samples due to high sensitivity of PCR methods results in undue treatment of patients without any symptoms of Pneumocystis pneumonia. The aim of the present study is to demonstrate Pneumocystis carinii in rats, immune suppressed by oral and subcutaneous administration of dexamethasone.

MATERIAL AND METHODS: Blood, oral, nasal and eye swabs were collected prior to immune suppression and 2, 6, 12 weeks after administration of dexamethasone. Also, samples were collected from lung, heart, liver, kidney, diaphragm, brain, spleen, tongue, muscle, eye, intestine, and feces. Cysts and trophozoites were investigated in stained slides and MSG gene was detected by PCR.

RESULTS: The results showed that weight loss is significantly higher in rats administered oral dexamethasone (P<0.05). Microscopy was positive only in lungs of rats orally administered dexamethasone. PCR was positive in lungs and oral swabs of rats prior to the administration of dexamethasone. After the administration of dexamethasone, the MSG gene was detected in oral swabs, lungs, spleen, kidney and (for the first time) in nasal swabs. PCR was positive in nasal swabs during the second and sixth weeks of oral and subcutaneous administration of dexamethasone, respectively.

CONCLUSIONS: Presence of P. jiroveci in nasopharyngeal aspirate, oropharyngeal wash, oral swab, induced sputum or BAL, and absence in nasal swab in a patient without symptoms of PCP may support clinician’s decision regarding colonization. Overall, detection of P. carinii in nasal swabs of rats by PCR demonstrated that nasal sampling can be used for the diagnosis of Pneumocystis pneumonia.

Keywords: Pneumonia, Pneumocystis - pathology, Pneumocystis jirovecii - isolation & purification, Nose - microbiology, Microscopy - methods, Immunosuppression, Genes, Fungal - genetics, Polymerase Chain Reaction - methods

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Medical Science Monitor Basic Research eISSN: 2325-4416
Medical Science Monitor Basic Research eISSN: 2325-4416